I measured the DNA concentration using Nanodrop 2000 but had no results.
- The DNA was extracted with a Norgen Biotek plant/fundi DNA isolation kit.
- All the procedures were followed as requested in the procedure checklist.
- Nanodrop was cleaned
We made 6 extractions from Silica gel preserved or fresh material and had the same results. Contaminants are present and basically no DNA was recovered.
Does somebody have any idea what could be wrong?
1. Lysis buffer expired?
2. RNAse A expired?
If somebody could provide a solution, it would be highly appreciated.
Problem with the starting material? If you still have the extract and you expect it to be a large amount of DNA (>10ng/ul), you may want to load 5ul to an agarose gel to see whether it is not an issue with Nanodrop
The Nanodrop is working. Two students are using it with perfect results (they are not using the Norgen kit for extraction, but a different protocol)
Issue could be with the starting materials, the kit, or the choice of protocol.
Have you (or anyone else) had success with this type kit before, even if it was for different starting materials? Do you have any reason to suspect the kit components are expired?
How did you process the starting material? If the sample is fibrous or dry, the classic method is to flash freeze in liquid nitrogen & use a mortar & pestle to create a fine powder. Or use a "bead beater" to homogenize in the starting buffer.
Does the starting material have a lot of lipids/oil or secondary metabolites? Sometimes a phenol chloroform extraction is necessary to remove those types of molecules.
I hope you can get this working soon!
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